Human DPF2 knockout A-431 cell line
- Advanced Validation
- What is this?
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- WB
Lab
Western blot - Human DPF2 knockout A-431 cell line (AB270473)
Lanes 1 - 4 : Merged signal (red and green). Green - ab134942 observed at 50 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab134942 was shown to react with DPF2/REQ in wild-type A-431 cells in western blot Loss of signal was observed when DPF2 knockout cell line ab270473 (knockout cell lysate ab270496) was used. Wild-type and DPF2 knockout A-431 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab134942 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-DPF2/REQ antibody [EPR9206(B)] (<a href='/en-us/products/primary-antibodies/dpf2-req-antibody-epr9206b-ab134942'>ab134942</a>) at 1/1000 dilution
Lane 1:
Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
DPF2 knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg
Lane 2:
Western blot - Human DPF2 knockout A-431 cell line (ab270473)
Lane 3:
HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4:
LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg
Predicted band size: 44 kDa
Observed band size: 50 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human DPF2 knockout A-431 cell line (AB270473)
1 bp deletion after Glu88 of the WT protein
- NGS
Supplier Data
Next Generation Sequencing - Human DPF2 knockout A-431 cell line (AB270473)
Knockout achieved by CRISPR/Cas9; X = 1 bp deletion; Frameshift = 99.77%
Complementary Products
Primary Antibodies
AB134942
Anti-DPF2/REQ antibody [EPR9206(B)]
RabMAb|Recombinant|KO Validated|Trial Size
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DPF2/REQ antibody [EPR9206(B)] (AB134942)](https://content.abcam.com/products/images/dpf2-req-antibody-epr9206b-ab134942--immunohistochemistry-formalin-pfa-fixed-paraffin-embedded-sections-img66288.jpg)
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Reactivity data
Product details
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant patents please refer to our limited use license and patent pages.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Handling procedures
Initial handling guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
DMEM (High Glucose) + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
DPF2 acts as a component of the BAF complex which is a multi-subunit chromatin remodeling complex. The BAF complex modifies the structure of chromatin to regulate gene transcription affecting gene expression outcomes. DPF2 helps recruit other important proteins to the BAF complex contributing to the control of gene transcription dynamics during cell differentiation and development. Its function is essential in maintaining proper chromatin state which is important for cellular identity and function.
Pathways
DPF2 interacts significantly within the Wnt and Notch signaling pathways. In the Wnt pathway DPF2 contributes to the regulation of genes involved in cell proliferation differentiation and apoptosis. This protein also interfaces with related pathways by influencing the activity of other proteins such as beta-catenin in the Wnt pathway. In the Notch signaling pathway DPF2 modulates gene expression by affecting chromatin structure which influences cellular processes like proliferation and differentiation.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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